The role of maternal caffeine consumption in the pathogenesis of adverse pregnancy outcomes is controversial. Several studies have found that women who consume caffeine are at increased risk of spontaneous abortion and fetal growth retardation compared to non-users. However, other equally well-done studies have found no harmful effects of caffeine consumption. In addition, several studies have reported that caffeine is harmful only among women who smoke. All previous studies of this question have relied on maternally-reported caffeine use; no studies have employed a biomarker for caffeine. This project first validated the use of serum caffeine and its metabolites as a marker for caffeine intake, and then studied these serum markers as a risk factor for adverse pregnancy outcome. In the validation study, serum paraxanthine was determined to be an acceptable marker for caffeine intake. As part of this project, the concentration of cotinine, a metabolite of nicotine, was assayed in the serum of approximately 450 of the women and the results compared to their reported cigarette smoking. Women were found to be very honestin reporting whether they smoked, but their serum concentration of cotinine correlated only moderately with the amount smoked. This was the case for two separate populations of women 30 years apart. In the main part of this project, the serum concentration of paraxanthine, caffeines primary metabolite, is being compared between 591 women experiencing a spontaneous abortion and 2558 women with live births who had serum drawn at the same time of pregnancy as the women with spontaneous loss. In addition, the association between reduced fetal growth and third-trimester paraxanthine serum concentrations will be evaluated among these 2558 women. The mean concentration of paraxanthine was higher in women experiencing a spontaneous abortion than women experiencing a live birth (752 vs 583 ng/ml). Further analysis revealed that this may be explained by a few women with very high concentrations of paraxanthine. The odds ratio for serum paraxanthine concentration greater than the 95th percentile was 1.86 (p<0.01), but intermediate concentrations were not associated with an elevated risk of spontaneous abortion. These results suggest that moderate caffeine consumption during pregnancy does not increase the risk of spontaneous abortion. - caffeine, spontaneous abortion, fetal growth, epidemiology, case-control study, biomarkers - Human Subjects: Interview, Questionaires, or Surveys Only